701317

antibody from Invitrogen Antibodies

Targeting: TIMP4

Western blot (Supportive data in Antibodypedia)

ELISA (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

701317

Provider

Invitrogen Antibodies

Product name

TIMP4 Recombinant Rabbit Monoclonal Antibody (13H4L13)

Antibody type

Monoclonal

Antigen

Other

Description

This antibody is predicted to react with bat, bovine, canine, equine, non-human primate, mouse, rabbit and rat based on sequence homology.

Antibody clone number

13H4L13

Concentration

0.5 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of TIMP4 was performed by loading 20 µg of Raji (lane1), HeLa (lane2), Jurkat (lane3) and MDA-MB-231 (lane4) cell lysates using Novex®NuPAGE®4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock Electrophoresis System (Product # EI0002), Novex® Sharp Pre-Stained Protein Standard (Product # LC5800), and iBlot® Dry Blotting System (Product # IB21001). Proteins were transferred to a nitrocellulose membrane and blocked with 5% skim milk for 1 hour at room temperature. TIMP4 was detected at ~25 kDa using TIMP4 Recombinant Rabbit Monoclonal Antibody (Product # 701317) at 1 µg-3 µg/mL in 2.5% skim milk at 4°C overnight on a rocking platform. Goat anti-Rabbit IgG-HRP Secondary Antibody (Product # G-21234) at 1:5000 dilution was used and chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of TIMP4 in whole cell extracts from MDA-MB-231 (30 µg) using a TIMP4 recombinant rabbit monoclonal antibody (Product # 701317) at a dilution of 2 µg/mL. Samples were detected using chemiluminescence (ECL). Results show a band at ~25 kDa.

ELISA

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Indirect ELISA analysis of endogenous TIMP4 in Hela lysate coated onto the plate using a TIMP4 recombinant rabbit monoclonal antibody (Product # 701317) at various dilutions. A non-linear regression analysis was performed (4 PL) and LOD and LOQ for the antibody were determined.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of TIMP4 in U2OS cells using a TIMP4 recombinant rabbit monoclonal antibody (Product # 701317) followed by detection using an Alexa Fluor 488-conjugated goat anti-rabbit secondary antibody (green) (Image A). Nuclei were stained using DAPI (Image B) and actin stained with Alexa Fluor 594 phalloidin (red) (image C). Image D is a composite image showing cytoplasmic localization of TIMP4.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry analysis of TIMP4 was done on HeLa cells. Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Tritonª X-100 for 20 minutes, and blocked with 5% BSA for 1 hour at room temperature. Cells were labeled with ABfinityª TIMP4 Recombinant Rabbit Monoclonal Antibody (701317, red histogram) or with rabbit isotype control (pink histogram) at 2 µg-4 µg/million cells in 2.5% BSA. After incubation at room temperature for 2-3 hours, the cells were labeled with Alexa Fluor¨ 488 Goat Anti-Rabbit Secondary Antibody (A11008) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10,000 cells were acquired and analyzed for each sample using an Attune¨ Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry analysis of TIMP4 in Hela cells using a TIMP4 recombinant rabbit monoclonal antibody (Product # 701317). Cells were fixed and permeabilized using FIX & PERM (Product # GAS-004) reagent, and detection was performed using an Alexa Fluor 488 goat anti-rabbit IgG (right peak) compared to an isotype control (left peak).

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Indirect ELISA analysis of endogenous TIMP4 in Hela lysate coated onto the plate using a TIMP4 recombinant rabbit monoclonal antibody (Product # 701317) at various dilutions. A non-linear regression analysis was performed (4 PL) and LOD and LOQ for the antibody were determined.